摘要
目的 活血软坚方的成药制剂-和络舒肝含药血清对肝星状细胞(HSC)-T6细胞株Smad3、Smad17、前胶原α2(Ⅰ)基因表达影响,以探讨活血软坚方抗肝纤维化作用及分子机制。方法 常规方法制备和络舒肝含药血清。用不同浓度和络舒肝含药血清干预HSC-T6细胞株,并以逆转录聚合酶链反应观察分析各组间Smad3、Smad7、前胶原α2(Ⅰ)mRNA表达的变化,以western blot检测Smad3蛋白的表达。结果 经和络舒肝含药血清处理,可下调HSC-T6细胞株Smad3、前胶原α2(Ⅰ) mRNA表达以及Smad3蛋白表达水平,前胶原α2(Ⅰ)表达变化与Smad3表达改变趋势一致。不同浓度的含药血清作用强度明显不同,药物作用后前胶原α2(Ⅰ) mRNA表达改变呈现血药浓度依赖性(F=6.92,P<0.05)。药物血清对Smad7 mRNA表达影响不明显(F=1.57,P>0.05)。结论 活血软坚方和络舒肝的抗肝纤维化作用可能是通过非特异性作用干扰转化生长因子β和Smad功能,抑制前胶原α2(Ⅰ) mRNA转录,从而减少细胞外基质生成。对Smad3而言,可能其表达升高产生的是促纤维化形成效应,反之,表现为抗纤维化作用。
Objective In order to explore the roles of Huoxueruanjian compound on liver fibrogenesis and its molecular mechanism, this paper has investigated the Influence of blood serum with such traditional Chinese medicine compound on the expression of Smad3, Smad7 and procollagen fα2 ( Ⅰ ) gene in hepatic stellate cell (HSC). Methods HSC-T6 was deal with different Concentration of blood serum medicine with Heluoshugan which was made by routine way. Then expression change of Smad3, Smad7 and procollagen α2 ( Ⅰ ) mRNA among each groups were observed by RT-PCR. Furthermore, the expression change of Smad3 protein were examined by Western blot. Results Expression of Smad3 and procollagen α2 ( Ⅰ ) mRNA as well as Smad3 protein had been downregulated after treating with blood serum medicine of Heluoshugan (P<0.01, P<0.05, respectively). The expression of procollagen α2 ( Ⅰ ) mRNA changed at the same tendency as those of Smad3. The role of blood serum medicine was significant difference between different concentration, P<0.05. And the expression of procollagen α2 ( Ⅰ ) mRNA changed in concentration-dependent manner. Blood serum medicine has no effects on the Smad7 mRNA. Conclusions The anti-fibrosis roles of HuoXueruanjian Compound maybe influence the function of TGF β and Smad by nonspecific action, thereby inhibit the transcription of procollegan α2 ( Ⅰ ) mRNA and decrease the production of ECM. As regards Smad3, it may be facilitating the development of liver fibrosis when its expression increases. Otherwise, it manifest with anti-fibrosis role.
出处
《中华肝脏病杂志》
CAS
CSCD
2004年第4期213-215,共3页
Chinese Journal of Hepatology
基金
广西自然科学基金(0135035)